Unexpected extensive lysine acetylation in the trump-card antibiotic producer Streptomyces roseosporus revealed by proteome-wide profiling
Unexpected extensive lysine acetylation in the trump-card antibiotic producer Streptomyces roseosporus revealed by proteome-wide profiling
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全蛋白质组分析揭示了王牌抗生素生产商玫瑰孢链霉菌中意外的广泛赖氨酸乙酰化
DOI：
10.1016/j.jprot.2014.04.017
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发表时间：
2014-06-25
影响因子：
3.3
通讯作者：
Xie, Jianping
中科院分区：
文献类型：
--
作者：
Liao, Guojian;Xie, Longxiang;Xie, Jianping
赖氨酸乙酰化是活细胞中普遍存在的一种保守的翻译后修饰。虽然赖氨酸乙酰化在调节初级代谢中的作用是公认的，但其在次级代谢中的功能在很大程度上仍然难以捉摸。为了深入了解被誉为“王牌抗生素”的达托霉素的生产者--疏孢链霉菌中赖氨酸乙酰化的性质、程度和生物学功能，我们采用基于免疫亲和的乙酰赖氨酸肽富集结合高分辨率质谱技术对该微生物中赖氨酸乙酰化蛋白进行了全面表征。我们共鉴定了667种蛋白质，具有1143个独特位点，代表了迄今为止在细菌中报道的最大的乙酰蛋白质组。根据基因本体论，乙酰化蛋白质属于各种功能类别，例如代谢和基因表达。我们第一次证明了参与不同次生代谢产物生物合成的蛋白质被乙酰化，如非核糖体肽合成酶，异羟肟酸铁载体和次膦酸天然产物生物合成所必需的酶，这意味着乙酰化在这些过程中的重要作用。总之，蛋白质组学分析揭示了赖氨酸乙酰化影响的细胞过程的惊人广度，也为抗生素生产者的合理改进提供了一些新的干预节点。生物学意义尽管已经付出了相当大的努力来阐明链霉菌中次级代谢的潜在机制，这是具有不同生物活性的次级代谢产物的多产者，如杀菌剂、抗肿瘤药物等，其监管全貌和相应网络还很不完善。蛋白质赖氨酸乙酰化是一种进化上保守的蛋白质翻译后修饰，广泛存在于各种生物学背景的蛋白质中。我们利用整合的高通量PTM蛋白质组学，然后进行深入的生物信息学分析，以分析玫瑰孢链霉菌的赖氨酸乙酰组。在667个赖氨酸乙酰化底物中，总共鉴定了1134个独特的Kac位点，代表了迄今为止原核生物中最大的乙酰组学。值得注意的是，非核糖体肽合成酶，异羟肟酸铁载体和次膦酸天然产物生物合成所必需的酶，被发现是乙酰化。鉴于这些酶在多种次生代谢产物的生物合成中的保守性，赖氨酸乙酰化可能在链霉菌的次生代谢调控中起重要作用。(C)2014爱思唯尔有限公司版权所有。
Lysine acetylation is emerging as a ubiquitous and conserved posttranslational modification in living cells. While the role of lysine acetylation in regulating primary metabolism is well-established, its function in secondary metabolism remains largely elusive. To gain insight into the nature, extent and biological function of lysine acetylation in Streptomyces reseosporus, a producer of daptomycin, dubiously dubbed as the trump card antibiotic, we used immunoaffinity-based acetyllysine peptide enrichment integrated with high resolution mass spectrometry to comprehensively characterize lysine acetylated proteins in this microbe. We identified a total of 667 proteins with 1143 unique sites, representing the largest acetylproteome reported to date in bacteria. Acetylated proteins belong to various functional classes such as metabolism and gene expression according to the gene ontology. We demonstrated for the first time that proteins involved in the biosynthesis of diverse secondary metabolites are acetylated, such as a nonribosomal peptide synthetase, enzymes essential for hydroxamate siderophore and phosphinic acid natural products biosynthesis, implying an important role of acetylation in these processes. Taken together, this proteomic analysis revealed a surprising breadth of cellular processes affected by lysine acetylation and also furnishes some fresh intervention nodes for the rational improvement of the antibiotic producer.Biological significanceDespite considerable efforts have been devoted to elucidating the mechanism underlying secondary metabolism in Streptomyces, which are prolific producers of secondary metabolites with diverse biological activities, such as bacteriocides and antitumors, the full map of regulation and corresponding network is still far from perfect. Protein lysine acetylation is an evolutionarily conserved protein post-translational modification, abundantly existing in proteins with diverse biological context. We took advantage of integrated high throughput PTM proteomics followed by intensive bioinformatic analysis to profile lysine acetylome of Streptomyces roseosporus. In total, 1134 unique Kac sites in 667 lysine acetylated substrates were identified, representing the largest aceylomics in prokaryotes to date. Significantly, a nonribosomal peptide synthetase, an enzyme essential for hydroxamate siderophore and phosphinic acid natural products biosynthesis, was found to be acetylated. Given the conservation of these enzymes in biosynthesis of diverse secondary metabolites, lysine acetylation likely plays an important role in regulating secondary metabolism in Streptomyces. (C) 2014 Elsevier B.V. All rights reserved.